Pig-a assay is a newly GLP-validated technique for measuring mutations caused by genotoxic effects of novel drug treatments or chemicals.
Gene mutation assays, like the Pig-a assay, measure the induction of mutations at a specific gene location as opposed to measuring overall DNA damage. This provides an advantage for product development and regulatory decision making.
Prior to the introduction of the Pig-a assay, other in vivo gene mutation assays had the disadvantages of being expensive and time-consuming, and required the use of specialized transgenic animals.
Clear Advantages of the Pig-a Assay include:
- Easily integrates into standard toxicity studies
- Reduces animal use
- Replaces expensive transgenic rodent mutation assays
- Fast, quantifiable results with flow cytometry
The entire Pig-a assay can be performed at BioReliance, or samples can be sent for analysis that can be performed alone or in conjunction with other endpoints (Micronucleus Assay, Comet Assay, Metaphase Analysis, etc.).
Consult with one of our gene mutation experts to determine if the Pig-a assay is the best approach for your project.
Pig-a Gene Mutation Assay Overview
The Pig-a Gene Mutation Assay is an in vivo assay primarily performed in rats, based on the X-linked Pig-a gene (phosphatidylinositol N-acetylglucosaminyltransferase, subunit A), which is involved in the production of glycosylohosphatidylino- sitol (GPI) anchor proteins on the cell surface.
The Pig-a gene is highly conserved in humans, rats, mice, and monkeys. Thus, this assay can help bridge the gap between preclinical models and humans. Single-step forward mutations in the Pig-a gene result in loss of the GPI anchored surface markers that can be detected early in immature red blood cells (reticulocytes) by flow cytometry.
Pig-a technology is relevant for both pharmaceutical development and chemical safety, and can easily be integrated into standard toxicity studies to provide additional and valuable data. This assay offers significant value as a secondary tier assay to understand a compound’s mode of action and to follow up a positive in vitro gene tox assay.
The Pig-a assay can be used to:
- Demonstrate mutations accumulated with repeat dosing
- Improve assay design and power
- Measure low dose responses and threshold effects
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